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https://locus.ufv.br//handle/123456789/12755
Tipo: | Artigo |
Título: | Trypanosoma cruzi nucleoside triphosphate diphosphohydrolase 1 (TcNTPDase-1) biochemical characterization, immunolocalization and possible role in host cell adhesion |
Autor(es): | Mariotini-Moura, Christiane Bastos, Matheus Silva e Castro, Felipe Freitas de Trindade, Mellina Lanna Vasconcellos, Raphael de Souza Neves-do-Valle, Myrian Augusta Araújo Moreira, Bernardo Pereira Santos, Ramon de Freitas Oliveira, Claudia Miranda de Cunha, Luana Celina Seraphim Souto, Xênia Macedo Bressan, Gustavo Costa Silva-Júnior, Abelardo Baqui, Munira Muhammad Abdel Bahia, Maria Terezinha Almeida, Márcia Rogéria de Meyer-Fernandes, José Roberto Fietto, Juliana Lopes Rangel |
Abstract: | Previous work has suggested that Trypanosoma cruzi diphosphohydrolase 1 (TcNTPDase-1) may be involved in the infection of mammalian cells and serve as a potential target for rational drug design. In this work, we produced recombinant TcNTPDase-1 and evaluated its nucleotidase activity, cellular localization and role in parasite adhesion to mammalian host cells. TcNTPDase-1 was able to utilize a broad range of triphosphate and diphosphate nucleosides. The enzyme's Km for ATP (0.096 mM) suggested a capability to influence the host's ATP-dependent purinergic signaling. The use of specific polyclonal antibodies allowed us to confirm the presence of TcNTPDase-1 at the surface of parasites by confocal and electron microscopy. In addition, electron microscopy revealed that TcNTPDase-1 was also found in the flagellum, flagellum insertion region, kinetoplast, nucleus and intracellular vesicles. The presence of this enzyme in the flagellum insertion region and vesicles suggests that it may have a role in nutrient acquisition, and the widespread distribution of TcNTPDase-1 within the parasite suggests that it may be involved in other biological process. Adhesion assays using anti-TcNTPDase-1 polyclonal antibodies as a blocker or purified recombinant TcNTPDase-1 as a competitor revealed that the enzyme has a role in parasite–host cell adhesion. These data open new frontiers to future studies on this specific parasite–host interaction and other unknown functions of TcNTPDase-1 related to its ubiquitous localization. |
Palavras-chave: | Recombinant protein Trypanosoma cruzi Nucleoside triphosphate Diphosphohydrolase Immunolocalization Adhesion |
Editor: | Acta Tropica |
Tipo de Acesso: | Open Access |
URI: | https://doi.org/10.1016/j.actatropica.2013.11.008 http://www.locus.ufv.br/handle/123456789/12755 |
Data do documento: | 19-Nov-2013 |
Aparece nas coleções: | Artigos |
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Arquivo | Descrição | Tamanho | Formato | |
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1-s2.0-S0001706X13003318-main.pdf | texto completo | 1,67 MB | Adobe PDF | Visualizar/Abrir |
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